白色念珠菌麦角甾醇通路影响变异链球菌致龋力的研究

王峥; 周学东; 任彪

doi:10.12182/20201160203

白色念珠菌麦角甾醇通路影响变异链球菌致龋力的研究

Ergosterol Pathway of Candida albicans Promotes the Growth and Cariogenic Virulence of Streptococcus mutans

摘要

摘要:
目的探究白色念珠菌（C. albicans）对变异链球菌（S. mutans）生长及毒力的影响，验证麦角甾醇通路在其中的作用。
方法将S. mutans菌株UA159、C. albicans野生型5314单独培养及共培养（体积比1∶1混合）24 h后，检测各组菌液光密度（OD）600 nm值，并利用菌落形成单位（colony-forming units，CFU）计数检测S. mutans浓度，以反映C. albicans对S. mutans生长的影响；利用蒽酮法检测生物膜胞外多糖产量，pH计检测菌液产酸能力，以反映C. albicans对S. mutans毒力的影响；将不同C. albicans麦角甾醇合成相关通路突变株与S. mutans共培养后，检测突变株对S. mutans生长的影响；利用0 mg/L、0.012 5 mg/L、0.025 mg/L氟康唑抑制C. albicans麦角甾醇合成相关通路，24 h后检测C. albicans野生型和S. mutans在单独培养和混合培养下CFU的变化，以验证该通路作用。
结果 C. albicans野生型与S. mutans共培养后，与S. mutans 单独培养组相比，混合菌液中OD₆₀₀值和CFU升高（P<0.05），S. mutans胞外多糖产量增加（P<0.05），pH下降更明显（P<0.05）。42株C. albicans突变株与S. mutans共培养后，有14株菌对S. mutans促进生长的作用消失（P<0.05），其中包含6株C. albicans麦角甾醇合成相关通路的突变株，这6株与S. mutans共培养后，菌液中S. mutans的CFU不变或下降。添加0.012 5 mg/L、0.025 mg/L氟康唑抑制麦角甾醇通路后，C. albicans 野生型与 S. mutans混合培养的菌液中，S. mutans的CFU较未添加氟康唑组降低（P<0.05），而C. albicans单独培养组、S. mutans单独培养组的CFU无明显变化（P>0.05），C. albicans野生型与S. mutans混合培养的菌液中C. albicans的CFU也无明显改变（P>0.05）。
结论 C. albicans能通过麦角甾醇相关通路增强S. mutans生长能力及毒力，此过程可被氟康唑抑制，有望成为防治龋病的新策略。

Abstract:
Objective To investigate the effect of Candida albicans (C. albicans) on proliferation and virulence of Streptococcus mutans (S. mutans), and to verify the role of the ergosterol pathway in it.
Methods After single- and co-cultivation of C. albicans wild-type 5314 and S. mutans UA159, the absorbance value of OD₆₀₀and colony-forming units (CFU) were detected to reflect the influence of C. albicans on the growth of S. mutans. To reflect the influence of C. albicans on the virulence of S. mutans, the production of extracellular polysaccharides was detected by anthrone-vitriol method, and the acid production capacity was detected by a pH meter. After single- and co-cultivation of C. albicans mutant strains and S. mutans, the growth of S. mutans was evaluated by CFU. After inhibiting the ergosterol pathway by 0 mg/L, 0.012 5 mg/L, and 0.025 mg/L fluconazole for 24 h, the CFU of single- and co-cultivated C. albicans wild-type and S. mutans were detected.
Results The OD₆₀₀absorbance value and CFU in theco-cultivation of C. albicans wild-type and S. mutans was higher than that in the single culture (P<0.05), and the production of extracellular polysaccharides in S. mutans was increased when S. mutans was co-cultured with C. albicans (P<0.05), accompanied with a more obvious decrease of pH (P<0.05). Fourteen strains in whole 42 C. albicans mutant strains lost the growth-promoting effect on S. mutans, including 6 ergosterol synthesis-related mutant strains. After co-cultivation of the 6 ergosterol synthesis-related mutant strains and S. mutans, the CFU of S. mutans remained unchanged or decreased. After inhibiting the ergosterol pathway by 0.012 5 mg/L and 0.025 mg/L fluconazole, the CFU of S. mutans in the co-cultivation of C. albicans wild-type and S. mutans was lower than that without fluconazole treatment (P<0.05), while the CFU of C. albicans and S. mutans single-cultivations did not change significantly (P>0.05) and the CFU of C. albicans in the co-cultivation of C. albicans wild-type and S. mutans did not change significantly (P>0.05).
Conclusion C.albicans can enhance the growth ability and virulence of S. mutans through the ergosterol-related pathway. This process can be inhibited by fluconazole, which is expected to become a novel strategy to prevent and treat dental caries.

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